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Bacillus amyloliquefaciens strain capable of controlling Ralstonia solanacearum effectively and promoting capsicum annuum growth

A technology for dissolving amyloid spores and bacilli, applied in the field of microorganisms, achieves obvious effects of promoting growth, promoting the growth of peppers, and having a wide antibacterial spectrum.

Active Publication Date: 2018-10-09
TOBACCO RES INST CHIN AGRI SCI ACAD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, there is no report about Bacillus amyloliquefaciens (Bacillus amyloliquefaciens) that can effectively prevent and control various pathogenic bacteria such as R. solani and can effectively promote the growth of pepper so far.

Method used

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  • Bacillus amyloliquefaciens strain capable of controlling Ralstonia solanacearum effectively and promoting capsicum annuum growth
  • Bacillus amyloliquefaciens strain capable of controlling Ralstonia solanacearum effectively and promoting capsicum annuum growth
  • Bacillus amyloliquefaciens strain capable of controlling Ralstonia solanacearum effectively and promoting capsicum annuum growth

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Experimental program
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Effect test

Embodiment 1

[0048] Embodiment 1: Determination of Bacillus amyloliquefaciens (Bacillus amyloliquefaciens) CAS02 antibacterial ability

[0049] The strain (CAS02) was isolated from tobacco rhizosphere soil in Qingdao, Shandong. The antibacterial ability of the strains against pathogenic bacteria was determined by the confrontation culture method (Sijam et al., 2005). Take 10 μL of bacterial suspension (OD420=0.3) and inoculate it in the center of NBY solid medium with a diameter of 9 cm. After the bacterial solution is fully absorbed, place the culture dish in a 28°C biochemical incubator and incubate it upside down for 72 hours. Then spray OD to NBY medium respectively 420 =0.3 pathogen suspension. The petri dish was transferred to a constant temperature incubator at 28°C again, and after upside-down cultivation for 24 hours, the antibacterial ability of the strain was measured. Xanthomonas campestris MSCT1, Ralstonia solanacearum, Pseudomonas syringae pv.tabaci and Colletotrichum acut...

Embodiment 2

[0050] Embodiment 2: the acquisition of strain 16S rDNA sequence

[0051] Using Bacillus amyloliquefaciens CAS02 genomic DNA as a template, the bacterial 16S rDNA universal primers 27F (5'-AGAGTTTGATCMTGGCTCAG-3') and 1492R (5'-TACGGYTACCTTGTTACGACTT-3') were used to amplify the 16S rDNA sequence. Amplification system and program: 25 μL PCR system contains 5.0 μL of 5×PCR buffer, 1.0 μL of DNA template (100 ng μL-1), 0.75 μL of dNTPs (10 mM), 0.75 μL of forward primer (10 μM), reverse primer ( 10μM) 0.75μL, MgCl2 (25mM) 2.5μL, Taq DNA polymerase (5UμL-1Promega) 0.25μL, and the rest was filled with ddH2O. The PCR amplification program was pre-denaturation at 94°C for 3 min; denaturation at 94°C for 30 s, annealing for 30 s, extension at 72°C for 1 min and 30 s, and 30 cycles. Finally, extend at 72°C for 10min and store at 4°C. After the reaction, the PCR products were analyzed by agarose gel electrophoresis. The obtained fragments were subjected to sequencing analysis. The ...

Embodiment 3

[0053] Embodiment 3: Growth-promoting test of bacterial strain CAS02 in greenhouse

[0054] Transplant pepper seedlings with consistent growth into the sterilized substrate, and when the seedlings grow to two true leaves, suspend and slowly pour CAS02 bacteria into the rhizome of pepper, 20ml per plant. The control group was irrigated with the same volume of Hoagland's nutrient solution. After 28 days, the growth-promoting effect of CAS02 on pepper was measured. Compared with the control, the treatment with CAS02 can significantly promote the growth of pepper plants, the fresh weight of the plants and the height of the above-ground parts increased by 81.25% and 34.23% respectively, and the chlorophyll a and chlorophyll b contents increased by 29.09% and 17.63% respectively compared with the control (Table 1, figure 2 ).

[0055] Table 1 The growth-promoting effect of strain CAS02 on pepper

[0056]

[0057]Note: Data are the average of three replicates. Different lett...

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Abstract

The invention provides a bacillus amyloliquefaciens strain capable of controlling Ralstonia solanacearum effectively and promoting capsicum annuum growth, and belongs to the technical field of microorganism. The bacillus amyloliquefaciens strain CAS02 is isolated from tobacco roots, is preserved at China General Microbiological Culture Collection Center on 26th, March, 2018, and the biological preservation number is CGMCC NO.15514. The bacillus amyloliquefaciens strain CAS02 is capable of promoting capsicum annuum growth, inhibiting growth of a plurality of pathogens such as capsicum annuum Pseudomonas solanacearum, and is promising in application prospect.

Description

technical field [0001] The invention belongs to the technical field of microorganisms, and in particular relates to a bacillus amyloliquefaciens that can effectively control R. solani and promote the growth of pepper. Background technique [0002] Pepper (Capsicum annuum L), belonging to Solanaceae, Capsicum genus, is an annual or limited perennial herb, and is an important vegetable crop and industrial raw material in my country. Bacterial wilt caused by Ralstonia solanacearum is a destructive soil-borne vascular disease on pepper, tomato, tobacco and other Solanaceous crops. The disease is widely distributed in tropical, subtropical and some temperate regions. With the continuous emergence of new hosts and the rapid development of facility agriculture, the occurrence of bacterial wilt is becoming more and more common and serious worldwide. At present, bacterial wilt has occurred and harmed in most provinces and autonomous regions of my country. According to different pla...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/20A01N63/02A01N63/00A01P1/00A01P21/00C12R1/07
CPCA01N63/00A01N63/10C12N1/20C12N1/205C12R2001/07
Inventor 王晓强王凤龙任广伟陈德鑫王静冯超王文静
Owner TOBACCO RES INST CHIN AGRI SCI ACAD
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